Proteases that degrade the amyloid [unreadable]-protein (A[unreadable]), which accumulates abnormally in Alzheimer's disease (AD), have emerged as critical regulators of amyloidogenesis in vivo, yet have only begun to be explored for their therapeutic potential. Accumulating experimental, genetic and animal modeling studies implicate insulin-degrading enzyme (IDE), as a particularly important A[unreadable]-degrading protease. Importantly, recent evidence shows how IDE activity might be increased by any of several mechanisms, including the displacement of endogenous inhibitors and modulation of its secretion into the extracellular space. Moreover, new crystal structures of IDE show that this protease possesses unorthodox enzymological properties that can be exploited to directly activate the protease as much as 40-fold. Here we propose to conduct ultra high- throughput screening (uHTS) on a chemically diverse library of ~550,000 compounds using a cell-based assay optimized for the detection of IDE activators. The development and implementation of the primary assay will largely be performed by Scripps Florida's highly experienced uHTS Core for a modest cost, permitting greater attention to be focused on critical secondary assays essential for identifying bone fide IDE activators and characterizing their mechanism(s) of action. Our long-term goal is to identify pharmacophores suitable for use in cultured cells and in vivo, which may lead to the development of novel therapies to treat this devastating disease. PUBLIC HEALTH RELELVANCE: The goal of this proposal is to test a large collection of molecules for their potential to affect fundamental biological processes known to regulate Alzheimer's disease, specifically relating to insulin-degrading enzyme. Discovered molecules will be evaluated for their possible therapeutic potential, and potentially developed into novel drugs through future funding proposals.